Methods of improving skin quality

ABSTRACT

Methods of improving skin quality are disclosed. Generally, the methods include topically administering an IRM compound to a treatment area of skin for a period of time and in an amount effective for improving the quality of the skin. Suitable IRM compound compounds include agonists of one or more TLRs.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority to U.S. Provisional Patent ApplicationNo. 60/454,245, filed Mar. 13, 2003.

BACKGROUND OF THE INVENTION

Skin condition is continuously affected by various factors including,for example, humidity, UV-light, cosmetics, aging, diseases, stress,cigarette smoking, and eating habits, each of which can result invarious skin changes. Additionally, certain changes appear on the skinthat are characteristic of aging, many of which are reflected, inparticular, by a change in the skin's structure. The main clinical signsof aging of the skin are, in particular, the appearance of fine linesand deep wrinkles, each of which can increase with age. Wrinkles can becaused by both the chronological aging of the skin and photoaging ofskin due to exposure of the skin to sunlight, UV-radiation and otherforms of actinic radiation.

In young skin, the collagen just beneath the surface of the skin formsan organized lattice with good elasticity and flexibility. During aging,the collagen structure can change, thus causing changes in the cosmeticappearance of the skin that many find undesirable. Current methods ofimproving skin quality include the application of cosmetic productscontaining active agents such as moisturizers, alpha-hydroxy acids,beta-hydroxy acids and retinoids. However, other methods are stillsought after.

SUMMARY OF THE INVENTION

It has been found that certain immune response modifier (IRM) compoundscan be used to improve skin quality.

In some aspects, the present invention provides methods of improvingskin quality by topically applying to the skin an IRM compound in anamount effective to improve the quality of the skin.

In some aspects, the present invention provides methods of visiblyreducing a skin change associated with aging by topically applying toskin exhibiting an age-associated change an IRM compound, wherein theIRM compound is applied in an amount and for a period of time sufficientto visibly reduce the skin change associated with aging.

In other aspects, the present invention includes methods of visiblyreducing a human skin wrinkle by topically applying to the human skinwrinkle an IRM compound in an amount and for a period of time sufficientto visibly reduce the wrinkle.

In other aspects, the present invention includes methods of treatingaging related skin conditions by topically applying to the skin an IRMcompound for a period of time and in an amount sufficient to effectchanges in the dermis.

In still other aspects, the present invention includes methods forreducing the appearance of skin changes associated with aging bytopically applying to an area of skin exhibiting skin changes associatedwith aging an IRM compound in an amount and for a period of timesufficient to reduce the appearance of skin changes associated withaging.

In yet other aspects, the present invention includes methods forimproving the quality of facial skin by topically applying to the facialskin an IRM compound in an amount and for a period of time sufficient toreverse or prevent changes in the dermis, where the changes in thedermis result from natural or innate aging or exposure to actinicradiation, and the changes in the dermis include diminution in thenumber and diameter of elastic fibers in the papillary dermis, atrophyof the dermis, reduction in subcutaneous adipose tissue, deposition ofabnormal elastic materials in the upper dermis, and combination thereof.

In some embodiments of the methods of the present invention, the IRMcompound may be an agonist of at least one TLR; including an agonist ofTLR7, TLR8 or both TR7 and TR8.

In some embodiments of the methods of the present invention, the IRMcompound may be administered via a topical application vehicle includinga cream, a foam, a gel, a spray, an ointment, a lotion, a solution, asuspension, a dispersion, an emulsion, a microemulsion, a paste, apowder, a wipe, or an oil.

In the some embodiments of the methods of the present invention, the IRMcompound may be an imidazoquinoline amine, a tetrahydroimidazoquinolineamine, an imidazopyridine amine, a 1,2-bridged imidazoquinoline amine, a6,7-fused cycloalkylimidazopyridine amine, an imidazonaphthyridineamine, a tetrahydroimidazonaphthyridine amine, an oxazoloquinolineamine, a thiazoloquinoline amine, an oxazolopyridine amine, athiazolopyridine amine, an oxazolonaphthyridine amine, or athiazolonaphthyridine amine, or a combination thereof.

Various other features and advantages of the present invention shouldbecome readily apparent with reference to the following detaileddescription, examples, and claims. In several places throughout thespecification, guidance is provided through lists of examples. In eachinstance, the recited list serves only as a representative group andshould not be interpreted as an exclusive list.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS OF THE INVENTION

The present invention provides methods of improving skin quality by thetopical administration of an immune response modifier (IRM) compound tothe skin in an amount effective to improve the quality of skin.

IRM compounds have previously been shown to be useful for treating manydifferent types of conditions. It has now been found that when topicallyapplied to the skin such as, for example, to treat a dermatologicalcondition, certain IRM compounds provide a secondary benefit ofimproving the quality of skin treated with the IRM. That is, treatmentof skin with an IRM compound not only, for example, clears lesionsassociated with the condition being treated, but also leaves the treatedskin in even better condition than skin unaffected by the condition and,therefore, left untreated.

IRM compounds include compounds that possess potent immunomodulatingactivity including but not limited to antiviral and antitumor activity.Certain IRMs modulate the production and secretion of cytokines. Forexample, certain IRM compounds induce the production and secretion ofcytokines such as, e.g., Type I interferons, TNF-α, IL-1, IL-6, IL-8,IL-10, IL-12, MIP-1, and/or MCP-1. As another example, certain IRMcompounds can inhibit production and secretion of certain T_(H)2cytokines, such as IL-4 and IL-5. Additionally, some IRM compounds aresaid to suppress IL-1 and TNF (U.S. Pat. No. 6,518,265).

Certain IRMs are small organic molecules (e.g., molecular weight underabout 1000 Daltons, preferably under about 500 Daltons, as opposed tolarge biological molecules such as proteins, peptides, and the like)such as those disclosed in, for example, U.S. Pat. Nos. 4,689,338;4,929,624; 4,988,815; 5,037,986; 5,175,296; 5,238,944; 5,266,575;5,268,376; 5,346,905; 5,352,784; 5,367,076; 5,389,640; 5,395,937;5,446,153; 5,482,936; 5,693,811; 5,741,908; 5,756,747; 5,939,090;6,039,969; 6,083,505; 6,110,929; 6,194,425; 6,245,776; 6,331,539;6,376,669; 6,451,810; 6,525,064; 6,541,485; 6,545,016; 6,545,017;6,558,951; 6,573,273; 6,656,938; 6,660,735; 6,660,747; 6,664,260;6,664,264; 6,664,265; 6,667,312; 6,670,372; 6,677,347; 6,677,348;6,677,349; 6,683,088; European Patent 0 394 026; U.S. Patent PublicationNos. 2002/0016332; 2002/0055517; 2002/0110840; 2003/0133913;2003/0199538; and 2004/0014779; and International Patent PublicationNos. WO 01/74343; WO 02/46749 WO 02/102377; WO 03/020889; WO 03/043572;WO 03/045391; and WO 03/103584.

Additional examples of small molecule IRMs include certain purinederivatives (such as those described in U.S. Pat. Nos. 6,376,501, and6,028,076), certain imidazoquinoline amide derivatives (such as thosedescribed in U.S. Pat. No. 6,069,149), certain imidazopyridinederivatives (such as those described in U.S. Pat. No. 6,518,265),certain benzimidazole derivatives (such as those described in U.S. Pat.No. 6,387,938), certain derivatives of a 4-aminopyrimidine fused to afive membered nitrogen containing heterocyclic ring (such as adeninederivatives described in U.S. Pat. Nos. 6,376,501; 6,028,076 and6,329,381; and in WO 02/08595), and certain3-β-D-ribofuranosylthiazolo[4,5-d]pyrimidine derivatives (such as thosedescribed in U.S. Publication No. 2003/0199461).

Other IRMs include large biological molecules such as oligonucleotide.sequences. Some IRM oligonucleotide sequences contain cytosine-guaninedinucleotides (CpG) and are described, for example, in U.S. Pat. Nos.6,194,388; 6,207,646; 6,239,116; 6,339,068; and 6,406,705. SomeCpG-containing oligonucleotides can include synthetic immunomodulatorystructural motifs such as those described, for example, in U.S. Pat.Nos. 6,426,334 and 6,476,000. Other IRM nucleotide sequences lack CpGsequences and are described, for example, in International PatentPublication No. WO 00/75304.

Other IRMs include biological molecules such as aminoalkyl glucosaminidephosphates (AGPs) and are described, for example, in U.S. Pat. Nos.6,113,918; 6,303,347; 6,525,028; and 6,649,172.

Certain IRMs can function as Toll-like receptor (TLR) agonists, i.e.,their immunomodulating influence is exerted through a TLR-mediatedcellular pathway. For example, some small molecule IRMs have beenidentified as agonists of one or more members of the TLR receptorfamily, TLR2, TLR4, TLR6, TLR7, and TLR8; certain AGPS have beenidentified as agonists of TLR4; and, certain CpGs have been identifiedas a agonists of TLR9. In many cases, activating a TLR-mediated pathwayresults in gene transcription, cytokine or co-stimulatory markerexpression regardless of the particular TLR that is activated.

In certain embodiments of the present invention, the IRM is an agonistof at least one TLR. In particular embodiments, the IRM compound can bean agonist of TLR7, TLR8, and/or TLR9. In alternative embodiments, theIRM compound is an agonist of TLR4. In certain specific embodiments, theIRM is an agonist of TLR8 or an agonist of both TLR7 and TLR8. The IRMmay induce the production of one or more cytokines, including but notlimited, to Type I interferons, TNF-α, IL-10, and IL-12. See, forexample, Gibson et al., Cell Immunol. 218(1-2):74-86 (2002). The IRM mayeffect the maturation, activation, and/or migration of cells of themyeloid lineage, including, but not limited to, macrophages, dendriticcells, and Langerhans cells.

Suitable IRM compounds include, but are not limited to, the smallmolecule IRM compounds described above having a 2-aminopyridine fused toa five membered nitrogen-containing heterocyclic ring. Such compoundsinclude, for example, imidazoquinoline amines including but not limitedto amide substituted imidazoquinoline amines, sulfonamide substitutedimidazoquinoline amines, urea substituted imidazoquinoline amines, arylether substituted imidazoquinoline amines, heterocyclic ethersubstituted imidazoquinoline amines, amido ether substitutedimidazoquinoline amines, sulfonamido ether substituted imidazoquinolineamines, urea substituted imidazoquinoline ethers, thioether substitutedimidazoquinoline amines, and 6-, 7-, 8-, or 9-aryl or heteroarylsubstituted imidazoquinoline amines; tetrahydroimidazoquinoline aminesincluding but not limited to amide substitutedtetrahydroimidazoquinoline amines, sulfonamide substitutedtetrahydroimidazoquinoline amines, urea substitutedtetrahydroimidazoquinoline amines, aryl ether substitutedtetrahydroimidazoquinoline amines, heterocyclic ether substitutedtetrahydroimidazoquinoline amines, amido ether substitutedtetrahydroimidazoquinoline amines, sulfonamido ether substitutedtetrahydroimidazoquinoline amines, urea substitutedtetrahydroimidazoquinoline ethers, and thioether substitutedtetrahydroimidazoquinoline amines; imidazopyridine amines including butnot limited to amide substituted imidazopyridine amines, sulfonamidosubstituted imidazopyridine amines, urea substituted imidazopyridineamines, aryl ether substituted imidazopyridine amines, heterocyclicether substituted imidazopyridine amines, amido ether substitutedimidazopyridine amines, sulfonamido ether substituted imidazopyridineamines, urea substituted imidazopyridine ethers, and thioethersubstituted imidazopyridine amines; 1,2-bridged imidazoquinoline amines;6,7-fused cycloalkylimidazopyridine amines; imidazonaphthyridine amines;tetrahydroimidazonaphthyridine amines; oxazoloquinoline amines;thiazoloquinoline amines; oxazolopyridine amines; thiazolopyridineamines; oxazolonaphthyridine amines; thiazolonaphthyridine amines; and1H-imidazo dimers fused to pyridine amines, quinoline amines,tetrahydroquinoline amines, naphthyridine amines, ortetrahydronaphthyridine amines. Various combinations of IRMs can be usedif desired.

In some embodiments, the IRM compound is an imidazoquinoline amine suchas, for example, 1-(2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-amine or4-amino-α,α-dimethyl-2-ethoxymethyl-1H-imidazo[4,5-c]quinolin-1-ethanol.In one particular embodiment, the IRM compound is1-(2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-amine.

In an alternative embodiment, the IRM compound is animidazonaphthyridine amine such as, for example,2-methyl-1-(2-methylpropyl)-1H-imidazo[4,5-c][1,5]naphthyridin-4-amineor 1-(2-methylpropyl)-1H-imidazo[4,5-c][1,5]naphthyridin-4-amine.

In another alternative embodiment, the IRM compound is a sulfonamidesubstituted imidazoquinoline amine such as, for example,N-[4-(4-amino-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl]methanesulfonamide.

In another alternative embodiment, the IRM compound is an amidesubstituted imidazoquinoline amine such as, for example,N-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]-1,1-dimethylethyl}cyclohexanecarboxamide.

In another alternative embodiment, the IRM compound is a thioethersubstituted imidazoquinoline amine such as, for example,2-butyl-1-[2-(propylsulfonyl)ethyl]-1H-imidazo[4,5-c]quinolin-4-amine.

In yet another alternative embodiment, the IRM compound is animidazopyridine amine such as, for example,N-{2-[4-amino-2-(ethoxymethyl)-6,7-dimethyl-1H-imidazo[4,5-c]pyridin-1-yl]ethyl}benzamide.

In certain embodiments, the IRM compound may be an imidazonaphthyridineamine, a tetrahydroimidazonaphthyridine amine, an oxazoloquinolineamine, a thiazoloquinoline amine, an oxazolopyridine amine, athiazolopyridine amine, an oxazolonaphthyridine amine, or athiazolonaphthyridine amine.

In certain embodiments, the IRM compound may be a substitutedimidazoquinoline amine, a tetrahydroimidazoquinoline amine, animidazopyridine amine, a 1,2-bridged imidazoquinoline amine, a 6,7-fusedcycloalkylimidazopyridine amine, an imidazonaphthyridine amine, atetrahydroimidazonaphthyridine amine, an oxazoloquinoline amine, athiazoloquinoline amine, an oxazolopyridine amine, a thiazolopyridineamine, an oxazolonaphthyridine amine, or a thiazolonaphthyridine amine.

As used herein, a substituted imidazoquinoline amine refers to an amidesubstituted imidazoquinoline amine, a sulfonamide substitutedimidazoquinoline amine, a urea substituted imidazoquinoline amine, anaryl ether substituted imidazoquinoline amine, a heterocyclic ethersubstituted imidazoquinoline amine, an amido ether substitutedimidazoquinoline amine, a sulfonamido ether substituted imidazoquinolineamine, a urea substituted imidazoquinoline ether, a thioethersubstituted imidazoquinoline amine, or a 6-, 7-, 8-, or 9-aryl orheteroaryl substituted imidazoquinoline amine. As used herein,substituted imidazoquinoline amines specifically and expressly exclude1-(2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-amine and4-amino-α,α-dimethyl-2-ethoxymethyl-1H-imidazo[4,5-c]quinolin-1-ethanol.

Unless otherwise indicated, reference to a compound can include thecompound in any pharmaceutically acceptable form, including any isomer(e.g., diastereomer or enantiomer), salt, solvate, polymorph, and thelike. In particular, if a compound is optically active, reference to thecompound can include each of the compound's enantiomers as well asracemic mixtures of the enantiomers.

An IRM compound may be provided in any formulation suitable foradministration to a subject. Suitable types of formulations aredescribed, for example, in U.S. Pat. No. 5,736,553; U.S. Pat. No.5,238,944; U.S. Pat. No. 5,939,090; U.S. Pat. No. 6,365,166; U.S. Pat.No. 6,245,776; U.S. Pat. No. 6,486,186; European Patent No. EP 0 394026; and U.S. Patent Publication No. 2003/0199538. The compound may beprovided in any suitable form including but not limited to a solution, asuspension, an emulsion, or any form of mixture. The compound may bedelivered in formulation with any pharmaceutically acceptable excipient,carrier, or vehicle. For example, the formulation may be delivered in aconventional topical dosage form such as, for example, a cream, anointment, an aerosol formulation, a non-aerosol spray, a gel, a foam, asolution, a suspension, a dispersion, an emulsion, a microemulsion, apaste, a powder, a solid stick (e.g., wax- or petroleum-based sticks), awipe, an oil, a lotion, and the like. In one particular embodiment, theIRM compound is provided in a cream formulation suitable for topicaladministration.

A formulation suitable for practicing the invention may include one ormore additional active ingredients such as, for example, another IRMcompound, acyclovir, valcyclovir, pencyclovir, amphotericins,chlorhexidine, clotrimazole, ketoconazole, econazole, miconazole,metronidazole, minocycline, nystatin, neomycin, kanamycin, phenytoin,octyl dimethyl PABA, octyl methoxycinnamate, PABA and other esters,octyl salicylate, oxybenzone, dioxybenzone, tocopherol, tocopherylacetate, selenium sulfide, zinc pyrithione, soluble elastin,diphenhydramine, pramoxine, lidocaine, procaine, erythromycin,tetracycline, clindamycin, crotamiton, hydroquinone and its monomethyland benzyl ethers, naproxen, ibuprofen, cromolyn, retinoic acid,retinol, retinyl palmitate, retinyl acetate, coal tar, griseofulvin,hydrocortisone, hydrocortisone 21-acetate, hydrocortisone 17-valerate,hydrocortisone 17-butyrate, betamethasone valerate, betamethasonedipropionate, triamcinolone acetonide, fluocinonide, clobetasolpropionate, minoxidil, dipyridamole, diphenylhydantoin, benzoylperoxide, 5-fluorouracil, vitamin A acetate (retinyl acetate) andvitamin E acetate (tocopheryl acetate). In some embodiments, additionalbeneficial effects may also be found when a skin-bleaching agent, suchas a hydroquinone (including glycolic acid, lactic acid, methyllacticacid, mandelic acid, pyruvic acid, methyl pyruvate, ethyl pyruvate,benzilic acid, gluconolactone, malic acid, tartaric acid, citric acid,and tropic acid) or a monobenzone, is incorporated into an IRMcomposition. In some embodiments, the IRM compound may be incorporatedinto, for example, a sunscreen, a skin lotion, a skin moisturizer, orother cosmetic.

The composition of a suitable formulation may depend at least in part onmany factors known in the art including but not limited to the physicaland chemical nature of the IRM compound; the nature of the carrier; thedosing regimen; the state of the subject's immune system (e.g.,suppressed, compromised, stimulated); the method of administering theIRM compound; and the desired result (e.g., wrinkle reduction,hydration, scar prevention, etc.). Accordingly it is not practical toset forth generally a single formulation suitable for improving skinquality for all possible applications. Those of ordinary skill in theart, however, can readily determine a suitable formulation with dueconsideration of such factors.

A suitable formulation may contain, for example, about 0.001%, about0.002%, about 0.005%, about 0.01%, about 0.015%, about 0.02%, about0.025%, about 0.05%, about 0.1%, about 0.25%, about 0.5%, about 0.75%,about 1%, about 2.5%, about 5%, about 7.5%, about 10%, about 25%, orabout 50% active IRM compound. In one particular embodiment, thecomposition includes about 5% IRM compound.

Skin treated by practicing the invention can include facial skin, skinon the neck, hands, arms, legs, or torso, and skin of other bodyregions.

Improving skin quality includes reversing, slowing the progression of,or preventing skin changes associated with natural or innate aging. Asused herein, “prevent” and variations thereof refer to any degree ofdelaying the onset of skin changes. For example, improving skin qualityincludes the reversal, slowing the progression of, or prevention of skinchanges associated with sun damage or photoaging—i.e., skin changesassociated with exposure to sunlight or other forms of actinic radiation(for example, such as UV radiation and tanning booths). As anotherexample, improving skin quality also can include reversing, slowing theprogression of, or preventing skin changes resulting from extrinsicfactors, including, but not limited to, radiation, air pollution, wind,cold, dampness, heat, chemicals, smoke, cigarette smoking, andcombinations thereof.

Improving skin quality also can include reversing, preventing orreducing scarring the can result, for example, from certain skinconditions (e.g., acne), infections (i.e., leishmaniasis), or injury(e.g., abrasions, punctures, lacerations, or surgical wounds).

Skin changes treatable by practicing the invention include, for example,wrinkles (including, but not limited to, human facial wrinkles),deepening of skin lines, thinning of skin, reduced scarring, yellowingof the skin, mottling, hyperpigmentation, appearance of pigmented and/ornon-pigmented age spots, leatheriness, loss of elasticity, loss ofrecoilability, loss of collagen fibers, abnormal changes in the elasticfibers, deterioration of small blood vessels of the dermis, formation ofspider veins, and combinations thereof.

Skin changes in the dermis also can be treated by practicing theinvention. Such changes in the dermis include, but are not limited to, areduction in the number and diameter of elastic fibers in the papillarydermis, atrophy of the dermis, reduction in subcutaneous adipose tissue,deposition of abnormal elastic materials in the upper dermis, andcombinations thereof.

Improving skin quality includes decreasing, reducing, and/or minimizingone or more of the skin changes discussed above. Improving skin qualitymay result in the skin having a more youthful appearance. Improving skinquality may result in the skin having a smoother, hydrated (i.e., lessdry), or less scaly appearance.

In some embodiments, an IRM compound may be administered to treat—i.e.,reverse or slow the progression of—one or more skin changes. Thus, theIRM compound may be administered after one or more skin changes haveoccurred. In other embodiments, an IRM compound may be administered toprevent one or more skin changes. Thus, the IRM compound may beadministered before one or more skin changes have occurred, to preventor slow the onset of such skin changes.

For example, in certain embodiments, improving skin quality can includea reduction in roughness, dryness, or scaliness. Skin qualityassessments, performed in conjunction with efficacy trials in whichcancerous (basal cell carcinoma, BCC) or pre-cancerous (actinickeratosis, AK) dermal lesions were treated with an IRM compound (5%imiquimod cream ALDARA, 3M Pharmaceuticals) indicate that treatment withthe IRM compound not only cleared the lesions, but also improved skinquality of the treated area.

The IRM compound was administered once daily either 5× per week or 7×per week for six weeks for treating BCC. Subjects in each treatmentgroup completed both an initiation (prior to the 6-week treatmentperiod) skin surface assessment and a follow-up (twelve weeks aftercompletion of the treatment period) skin surface assessment. Skinquality was assessed on a scale of 1 (none) to 4 (severe). Theassessment from the initiation visit established a baseline againstwhich the follow-up assessment was compared. Both of the IRM-treatedgroups (5× per week and 7× per week) showed a substantial decrease inthe degree of rough/dry/scaly skin surface over the treatment area. Theresults (see Table 1) were statistically significant over the baselineas well as statistically significant over the placebo-treated controlgroup.

In a separate study, the IRM was administered once daily either 2× perweek or 3× per week for sixteen weeks for treating AK. Subjects in eachtreatment group completed both an initiation (prior to the 16-weektreatment period) skin surface assessment and a follow-up (eight weeksafter completion of the treatment period) skin surface assessment. Skinquality was assessed on a scale of 1 (none) to 4 (severe). Theassessment from the initiation visit established a baseline againstwhich the follow-up assessment was compared. Both of the IRM-treatedgroups (2× per week and 3× per week) showed a substantial decrease inthe degree of rough/dry/scaly skin surface over the treatment area. Theresults (see Table 2) for both groups were statistically significantagainst both the baseline assessments and the placebo-treated controlgroup.

The particular amount of IRM compound necessary to improve skin qualitymay depend, at least in part, on one or more factors. Such factorsinclude, but are not limited to, the particular IRM compound beingadministered; the state of the subject's overall health; the state ofthe subject's immune system (e.g., suppressed, compromised, stimulated);the route of administering the IRM; and the desired result (e.g.,wrinkle reduction, reducing dryness, etc.). Accordingly, it is notpractical to set forth generally the amount that constitutes an amountof an IRM compound effective for improving skin quality. Those ofordinary skill in the art, however, can readily determine theappropriate amount with due consideration of such factors.

In some embodiments, the methods of the present invention includeadministering sufficient IRM compound to provide a dose of, for example,from about 100 ng/kg to about 50 mg/kg to the subject, although in someembodiments the method may be performed by administering IRM compound ina dose outside this range. In some of these embodiments, the methodincludes administering sufficient IRM compound to provide a dose of fromabout 10 μg/kg to about 5 mg/kg to the subject, for example, a dose offrom about 100 μg/kg to about 1 mg/kg.

The dosing regimen may depend at least in part on many factors known inthe art such as, for example, the physical and chemical nature of theIRM compound; the nature of the carrier; the amount of IRM beingadministered; the period over which the IRM compound is beingadministered; the state of the subject's immune system (e.g.,suppressed, compromised, stimulated); the method of administering theIRM compound; and the desired result. Accordingly it is not practical toset forth generally the dosing regimen effective for improving skinquality for all possible applications. Those of ordinary skill in theart, however, can readily determine an appropriate dosing regimen withdue consideration of such factors.

In some embodiments of the invention, the IRM compound may beadministered, for example, from a single dose to multiple dosesadministered multiple times per day. In certain embodiments, the IRMcompound may be administered from about once per week to about once perday. In one particular embodiment, the IRM compound is administered onceper day, two days per week. In an alternative embodiment, the IRMcompound is administered once per day, three times per week. In anotheralternative embodiment, the IRM compound is administered one per day,five days per week. In yet another alternative embodiment, the IRMcompound is administered once per day, seven days per week.

The period of time that is sufficient for practicing the invention maydepend, at least in part, on factors such as, for example, the physicaland chemical nature of the IRM compound; the nature of the carrier; theamount of IRM being administered; the frequency with which the IRMcompound is being administered; the state of the subject's immune system(e.g., suppressed, compromised, stimulated); the method of administeringthe IRM compound; and the desired result. Accordingly it is notpractical to set forth generally the period of time necessary to improveskin quality for all possible applications. Those of ordinary skill inthe art, however, can readily determine an appropriate period of timewith due consideration of such factors.

In some embodiments, a sufficient period of time may range from at leastone day to about six months, although in some embodiments the inventionmay be practiced by administering IRM compound for a period outside thisrange. In some embodiments, the IRM compound may be administered for atleast one week. In an alternative embodiment, the IRM compound may beadministered for at least about four weeks. In another alternativeembodiment, the IRM compound may be administered for at least abouteight weeks. In another alternative embodiment, the IRM compound may beadministered for at least about sixteen weeks.

The methods of the present invention may be performed on any suitablesubject. Suitable subjects include, but are not limited to, animals suchas, but not limited to, humans, non-human primates, rodents, dogs, cats,horses, pigs, sheep, goats, or cows.

EXAMPLES

The following examples have been selected merely to further illustratefeatures, advantages, and other details of the invention. It is to beexpressly understood, however, that while the examples serve thispurpose, the particular materials and amounts used as well as otherconditions and details are not to be construed in a matter that wouldunduly limit the scope of this invention. Unless otherwise indicated,all percentages and ratios are by weight.

Example 1 Treatment of Rough, Dry, or Scaly Skin

Volunteer subjects with superficial basal cell carcinoma (BCC) wererandomized to either the 5% imiquimod cream formulation (ALDARA, 3MPharmaceuticals, St. Paul, Minn.) or a placebo cream (Vehicle) in one oftwo treatment regimens: (1) once daily for seven days per week(7×/week), and (2) once daily for five consecutive days per week and notreatment for the remaining two days (5×/week). Subjects in each groupreceived treatment for six weeks.

Subjects were instructed to administer a single application of cream(Vehicle or 5% imiquimod, as assigned) to a target tumor just prior tonormal sleeping hours according to the dosing regimen to which they wereassigned. The subjects were instructed to wash the tumor lesion prior toapplying the cream, and then rub the cream into the tumor and intoextramarginal skin about 1 cm around the tumor. The subjects wereinstructed to leave the cream in place for at least eight hours withoutocclusion.

Subjects completed interval visits 1, 3, and 6 weeks after treatment wasinitiated and at twelve weeks after the end of treatment. At the12-weeks post-treatment visit, the treatment area was clinically andhistologically evaluated for evidence of BCC.

In addition, the treatment area was evaluated for skin quality. Skinquality was assessed on a scale of 1 (none) to 4 (severe). Theassessment from the Initiation visit established a baseline againstwhich the Follow-up assessment was compared. Both IRM-treated groups (5×per week and 7× per week) showed a substantial decrease in the degree ofrough/dry/scaly skin surface over the treatment area. The results werestatistically significant over the baseline as well as statisticallysignificant over the placebo-treated control group. Results are shown inTable 1.

TABLE 1 Treatment Visit N = None Mild Moderate Severe IRM 5x Initiation185 76 (41%) 93 (50%) 16 (9%)  0 (0%) Follow-up 178 141 (79%)  36 (20%)1 (1%) 0 (0%) Vehicle 5x Initiation 178 53 (30%) 105 (59%)  20 (11%) 0(0%) Follow-up 173 82 (47%) 82 (47%) 8 (5%) 1 (1%) IRM 7x Initiation 17966 (36%) 96 (54%) 16 (9%)  1 (1%) Follow-up 168 139 (83%)  27 (16%) 2(1%) 0 (0%) Vehicle 7x Initiation 181 56 (31%) 106 (59%)  19 (10%) 0(0%) Follow-up 169 88 (52%) 73 (43%) 7 (4%) 1 (1%)

Separately, volunteer subjects with actinic keratoses (AK) wererandomized to either the 5% imiquimod cream formulation (ALDARA, 3MPharmaceuticals, St. Paul, Minn.) or a placebo cream (Vehicle) in one oftwo treatment regimens: (1) once daily for two days per week (2×/week),and (2) once daily for three days per week (3×/week). Subjects in eachgroup received treatment for sixteen weeks.

Subjects were instructed to administer a single application of cream(Vehicle or 5% imiquimod, as assigned) to a 25 cm² treatment area justprior to normal sleeping hours according to the dosing regimen to whichthey were assigned. The subjects were instructed to wash the treatmentarea prior to applying the cream, and then rub the cream into thetreatment area. The subjects were instructed to leave the cream in placefor at least eight hours without occlusion.

Subjects completed interval throughout 16-week the treatment and ateight weeks after the end of treatment. At the 8-weeks post-treatmentvisit, the treatment area was clinically and histologically evaluatedfor evidence of AK.

In addition, the treatment area was evaluated for skin quality. Skinquality was assessed on a scale of 1 (none) to 4 (severe). Theassessment from the Initiation visit established a baseline againstwhich the Follow-up assessment was compared. Both IRM-treated groups (2×per week and 3× per week) showed a substantial decrease in the degree ofrough/dry/scaly skin surface over the treatment area. The results werestatistically significant over the baseline as well as statisticallysignificant over the placebo-treated control group. Results are shown inTable 2.

TABLE 2 Treatment Visit N = None Mild Moderate Severe IRM 2x Initiation215 26 (12.1%) 135 (62.8%) 50 (23.3%) 4 (1.9%) Follow-up 205 116(56.6%)   76 (37.1%) 13 (6.3%)  0 (0%)  Vehicle 2x Initiation 221 33(14.9%) 141 (63.8%) 44 (19.9%) 3 (1.4%) Follow-up 210 45 (21.4%) 123(58.6%) 38 (18.1%) 4 (1.9%) IRM 3x Initiation 242 43 (17.8%) 147 (60.7%)51 (21.1%) 1 (0.4%) Follow-up 226 132 (58.4%)   88 (38.9%) 6 (2.7%) 0(0%)  Vehicle 3x Initiation 250 46 (18.4%) 144 (57.6%) 57 (22.8%) 3(1.2%) Follow-up 233 58 (24.9%) 138 (59.2%) 36 (15.5%) 1 (0.4%)

Example 2

Subjects having cutaneous leishmaniasis received standard care forleishmaniasis: meglumine antimonate (GLUCANTIME, Aventis Pharma, 20mg/Kg) for 20 consecutive days. Subjects were randomized to received, inaddition to the meglumine antimonate, either 5% imiquimod cream (ALDARA,3M Pharmaceuticals, St. Paul, Minn.) or a placebo cream. A thin layer ofcream was applied to each lesion every other day for 20 days (i.e., tentotal applications). Doses of cream were applied by study personnelblinded to group assignments. Cream was applied with a gentle rubbingaction over areas of involved but intact skin—i.e., the whole area ofnodular lesions and including the periphery of ulcerative lesions (up to0.5 cm beyond the edge of each lesion).

Treatment efficacy was evaluated at follow-up visits one, two, three,six, and twelve months after the completion of treatment. Scar qualitywas not an original outcome of the study and thus, no well-standardizedscale for the assessment of scar quality was established. Nevertheless,study personnel blinded to group assignment throughout the treatmentperiod and follow-up visits were able, prior to unblinding, to identifywhich subjects had received imiquimod during the treatment period.

Example 3 Treatment of Wrinkles

Wrinkles of skin may be due to natural aging and/or sun damage. Mostfine wrinkles on the face are due to natural or innate aging, whilecoarse wrinkles on the face are the consequence of actinic or sundamage. Although the real mechanism of wrinkles formation in the skin isstill unknown, it has been shown that visible fine wrinkles are due todiminution in the number and diameter of elastic fibers in the papillarydermis, and also due to atrophy of dermis as well as reduction insubcutaneous adipose tissue. Histopathology and electron microscopystudies indicate that coarse wrinkles are due to excessive deposition ofabnormal elastic materials in the upper dermis and thickening of theskin.

A 5% cream of imiquimod, the imidazoquinoline amine1-(2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-amine, marketed as ALDARA(3M Pharmaceuticals, St. Paul, Minn.), is provided to a randomizedsegment of volunteer subjects having facial wrinkles lateral andinferior to the lateral canthus. The remaining subjects receive aplacebo formulation. The subjects are instructed to apply theformulation provided to them (ALDARA or placebo) at least daily on areasof facial wrinkles for 4 to 12 months. All subjects are told to avoidsun exposure, and to use sunscreen products if exposure to sunlight isunavoidable.

Evaluations are performed at the beginning of the study to establish abaseline, and at three month intervals during the study period.Evaluations include examination of the treatment area and photographingeach side the subject's face. The subjects are asked not to wear anyfacial make-up at the time of each photographic session. Standardizedphotographic conditions are used. At the end of the study, clinicalevaluations and review of photographs reveals substantial reductions infacial wrinkles. Clinical evaluations are performed using to thefollowing scale:

None (0): No evidence of wrinkling.

Mild (1): Minimal evidence of wrinkles beyond lateral canthus, wrinklesare fine and shallow.

Moderate (2): Superficial wrinkles that extend beyond orbital rim,wrinkles do not fold onto each other.

Severe (3): Deep folds that extend beyond orbital rim, wrinkles begin tofold onto each other.

Evaluation scores are analyzed to establish statistical significance ofchanges in evaluation scores (a) over the course of the study (baselinevs. end of study), and (b) with respect to placebo (ALDARA vs. placebo).The degree of improvement and reduction in wrinkles is evaluated anddetermined to range from mild improvement in some subjects, to verysubstantial improvement in other subjects.

Example 4 Treatment of Mottled Pigmentation

Many small and large discolored lesions such as, for example, age spots(solar lentigines) develop on the face and the back of the hands withaging.

A 5% cream of imiquimod, the imidazoquinoline amine1-(2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-amine, marketed as ALDARA(3M Pharmaceuticals, St. Paul, Minn.), is provided to a randomizedsegment of volunteer subjects having age spots and/or other pigmentedlesions. The remaining subjects receive a placebo formulation. Thesubjects are instructed to apply the formulations provided to them(ALDARA or placebo) to the age spots and/or other pigmented lesions.Specific instructions are given to the subjects that the medications areto be applied at least daily to the lesions of age spots and/or otherpigmented lesions.

Evaluations are performed at the beginning of the study to establish abaseline and throughout the study period. Evaluations includeexamination of the treatment area and photographing the treatment area.Standardized photographic conditions are used. At the end of the study,clinical evaluations and review of photographs reveals substantialreduction of irregular pigmentation. Clinical evaluations are performedusing to the following scale:

None (1): No evidence of irregular pigmentation changes.

Mild (2): Minimal evidence—in both extent and noticeability in contrastwith surrounding normal skin—of diffuse reticulated, irregularpigmentation changes, solar lentigines, or discretehypo/hyperpigmentated macules.

Moderate (3): Moderate evidence of one or more of the followingfindings: moderate diffuse reticulated, irregular pigmentation changes,solar lentigines, or discrete hypo/hyperpigmentated macules.

Severe (4): One or more of the following findings: Extensive reticulatedbackground irregular pigmentation changes, large discretehypo/hyperpigmentated macules, or solar lentigines.

Evaluation scores are analyzed to establish statistical significance ofchanges in evaluation scores (a) over the course of the study (baselinevs. end of study), and (b) with respect to placebo (ALDARA vs. placebo).At the end of 4 to 8 weeks, improvement of age spots is clinicallydiscernible. After 4 to 6 months of topical treatment, substantialimprovement of age spots is observed in the majority of subjects tested.Complete eradication of age spots is observed after 6 to 9 months oftopical administration with the IRM compositions of the instantinventions.

The complete disclosures of the patents, patent documents andpublications cited herein are incorporated by reference in theirentirety as if each were individually incorporated. In case of conflict,the present specification, including definitions, shall control. Variousmodifications and alterations to this invention will become apparent tothose skilled in the art without departing from the scope and spirit ofthis invention. Illustrative embodiments and examples are provided asexamples only and are not intended to limit the scope of the presentinvention. The scope of the invention is limited only by the claims setforth as follows.

What is claimed is:
 1. A method of visibly reducing a skin changeassociated with aging comprising: topically applying to skin exhibitinga change associated with aging an imiquimod composition comprising 5%imiquimod by weight in an amount and for a period of time sufficient tovisibly reduce the skin change associated with aging, wherein imiquimodis the sole active pharmaceutical ingredient applied to the human skinin performance of the method.
 2. A method of visibly reducing a humanskin wrinkle comprising: topically applying to the human skin wrinkle animiquimod composition comprising 5% imiquimod by weight in an amount andfor a period of time sufficient to visibly reduce the wrinkle.
 3. Themethod in accordance with claim 1, wherein the skin change is selectedfrom the group consisting of wrinkles, deepening of skin lines thinningof skin, reduced scarring, yellowing of the skin, mottling,hyperpigmentation, appearance of pigmented and/or non-pigmented agespots, leatheriness, loss of elasticity, loss of recoilability, loss ofcollagen fibers, abnormal changes in the elastic fibers, deteriorationof small blood vessels of the dermis, formation of spider veins, andcombinations thereof.
 4. The method in accordance with claim 3, whereinthe skin change is selected from the group consisting of yellowing ofthe skin, mottling, hyperpigmentation, and appearance of pigmentedand/or non-pigmented age spots.
 5. The method of claim 1, wherein theimiquimod composition is applied daily.
 6. The method of claim 1,wherein the imiquimod composition is applied once per day two to threetimes a week.
 7. The method of claim 1, wherein the imiquimodcomposition is applied to the skin exhibiting the change for about onemonth.
 8. The method of claim 1, wherein the imiquimod composition isapplied to the skin exhibiting the change for about two months.
 9. Themethod of claim 1, wherein the imiquimod composition is to the skinexhibiting the change for about 4 to 12 months.
 10. The method of claim1, further comprising measuring the reduction in the skin change byvisual or photographic examination of the skin.
 11. The method of claim1, wherein the imiquimod composition is applied to the human skinwrinkle three times a week.
 12. The method of claim 2, wherein theimiquimod composition is applied daily.
 13. The method of claim 2,wherein the imiquimod composition is applied once per day two to threetimes a week.
 14. The method of claim 2, wherein the imiquimodcomposition is applied to the human skin wrinkle for about one month.15. The method of claim 2, wherein the imiquimod composition is appliedto the human skin wrinkle for about two months.
 16. The method of claim2, wherein the imiquimod composition is applied to the human skinwrinkle for about 4 to 12 months.
 17. The method of claim 2, furthercomprising measuring the reduction in human skin wrinkles by visual orphotographic examination of the skin.
 18. The method of claim 2, whereinthe imiquimod composition is applied to the human skin wrinkle threetimes a week.